Muscarinic acetylcholine receptor antagonists

ABSTRACT

Muscarinic Acetylcholine Receptor Antagonists and methods of using them are provided.

FIELD OF THE INVENTION

This application is a continuation of U.S. application Ser. No.10/575,839, now U.S. Pat. No. 7,276,521, which is the §371 nationalstage entry of PCT/US2004/033638, filed 12 Oct. 2004 which claims thebenefit of priority of U.S. Provisional Application No. 60/511,009 filed14 Oct. 2003.

This invention relates to novel derivatives of8-azoniabicyclo[3,2,1]octanes, pharmaceutical compositions, processesfor their preparation, and use thereof in treating M₃ muscarinicacetylcholine receptor mediated diseases.

BACKGROUND OF THE INVENTION

Acetylcholine released from cholinergic neurons in the peripheral andcentral nervous systems affects many different biological processesthrough interaction with two major classes of acetylcholinereceptors—the nicotinic and the muscarinic acetylcholine receptors.Muscarinic acetylcholine receptors (mAChRs) belong to the superfamily ofG-protein coupled receptors that have seven transmembrane domains. Thereare five subtypes of mAChRs, termed M₁-M₅, and each is the product of adistinct gene. Each of these five subtypes displays uniquepharmacological properties. Muscarinic acetylcholine receptors arewidely distributed in vertebrate organs, and these receptors can mediateboth inhibitory and excitatory actions. For example, in smooth musclefound in the airways, bladder and gastrointestinal tract, M₃ mAChRsmediate contractile responses. For review, please see {Brown 1989247/id}.

Muscarinic acetylcholine receptor dysfunction has been noted in avariety of different pathophysiological states. For instance, in asthmaand chronic obstructive pulmonary disease (COPD), inflammatoryconditions lead to loss of inhibitory M₂ muscarinic acetylcholineautoreceptor function on parasympathetic nerves supplying the pulmonarysmooth muscle, causing increased acetylcholine release following vagalnerve stimulation. This mAChR dysfunction results in airwayhyperreactivity mediated by increased stimulation of M₃ mAChRs{Costello, Evans, et al. 1999 72/id} {Minette, Lammers, et al. 1989248/id}. Similarly, inflammation of the gastrointestinal tract ininflammatory bowel disease (IBD) results in M₃ mAChR-mediatedhypermotility {Oprins, Meijer, et al. 2000 245/id}. Incontinence due tobladder hypercontractility has also been demonstrated to be mediatedthrough increased stimulation of M₃ mAChRs {Hegde & Eglen 1999 251/id}.Thus the identification of subtype-selective mAChR antagonists may beuseful as therapeutics in these mAChR-mediated diseases.

Despite the large body of evidence supporting the use of anti-muscarinicreceptor therapy for treatment of a variety of disease states,relatively few anti-muscarinic compounds are in use in the clinic. Thus,there remains a need for novel compounds that are capable of causingblockade at M₃ mAChRs. Conditions associated with an increase instimulation of M₃ mAChRs, such as asthma, COPD, IBD and urinaryincontinence would benefit by compounds that are inhibitors of mAChRbinding.

SUMMARY OF THE INVENTION

This invention provides for a method of treating a muscarinicacetylcholine receptor (mAChR) mediated disease, wherein acetylcholinebinds to an M₃ mAChR and which method comprises administering aneffective amount of a compound of Formula (I) or Formula (II) [exceptthe compound of Formula (II) with R2 and R3 as 2-thiophene and R4 as—OC(O)CH₃] or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of inhibiting the binding ofacetylcholine to its receptors in a mammal in need thereof whichcomprises administering to aforementioned mammal an effective amount ofa compound of Formula (I) or Formula (II).

The present invention also provides for the novel compounds of Formula(I) or Formula (II), and pharmaceutical compositions comprising acompound of Formula (I) or Formula (II), and a pharmaceutical carrier ordiluent.

Compounds of Formula (I) or Formula (II) useful in the present inventionare represented by the structure:

wherein:

-   the H atom indicated is in the exo position;-   R1⁻ represents an anion associated with the positive charge of the N    atom. R1⁻ may be but is not limited to chloride, bromide, iodide,    sulfate, benzene sulfonate and toluene sulfonate;-   R2 and R3 are independently selected from the group consisting of    straight or branched chain lower alkyl groups (having preferably    from 1 to 6 carbon atoms), cycloalkyl groups (having from 5 to 6    carbon atoms), cycloalkyl-alkyl (having 6 to 10 carbon atoms),    heterocycloalkyl (having 5 to 6 carbon atoms) and N or O as the    heteroatom, heterocycloalkyl-alkyl (having 6 to 10 carbon atoms) and    N or O as the heteroatom, aryl, optionally substituted aryl,    heteroaryl, and optionally substituted heteroaryl;-   R4 is selected from the group consisting of (C₁-C₆)alkyl,    (C₃-C₁₂)cycloalkyl, (C₃-C₇)heterocycloalkyl,    (C₁-C₆)alkyl(C₃-C₁₂)cycloalkyl, (C₁-C₆)alkyl(C₃-C₇)heterocycloalkyl,    aryl, heteroaryl, (C₁-C₆)alkyl-aryl, (C₁-C₆)alkyl-heteroaryl, —OR5,    —CH₂OR5, —CH₂OH, —CN, —CF₃, —CH₂O(CO)R6, —CO₂R7, —CH₂NH₂,    —CH₂N(R7)SO₂R5, —SO₂N(R7)(R8), —CON(R7)(R8), —CH₂N(R8)CO(R6),    —CH₂N(R8)SO₂(R6), —CH₂N(R8)CO₂(R5), —CH₂N(R8)CONH(R7);-   R5 is selected from the group consisting of (C₁-C₆)alkyl,    (C₁-C₆)alkyl(C₃-C₁₂)cycloalkyl, (C₁-C₆)alkyl(C₃-C₇)heterocycloalkyl,    (C₁-C₆)alkyl-aryl, (C₁-C₆)alkyl-heteroaryl;-   R6 is selected from the group consisting of (C₁-C₆)alkyl,    (C₃-C₁₂)cycloalkyl, (C₃-C₇)heterocycloalkyl,    (C₁-C₆)alkyl(C₃-C₁₂)cycloalkyl, (C₁-C₆)alkyl(C₃-C₇)heterocycloalkyl,    aryl, heteroaryl, (C₁-C₆)alkyl-aryl, (C₁-C₆)alkyl-heteroaryl;-   R7 and R8 are, independently, selected from the group consisting of    H, (C₁-C₆)alkyl, (C₃-C₁₂)cycloalkyl, (C₃-C₇)heterocycloalkyl,    (C₁-C₆)alkyl(C₃-C₁₂)cycloalkyl, (C₁-C₆)alkyl(C₃-C₇)heterocycloalkyl,    (C₁-C₆)alkyl-aryl, and (C₁-C₆)alkyl-heteroaryl.

Suitable pharmaceutically acceptable salts are well known to thoseskilled in the art and include basic salts of inorganic and organicacids, such as hydrochloric acid, hydrobromic acid, sulphuric acid,phosphoric acid, methane sulphonic acid, ethane sulphonic acid, aceticacid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid and mandelic acid. In addition, pharmaceuticallyacceptable salts of compounds of Formula (I) or Formula (II) may also beformed with a pharmaceutically acceptable cation. Suitablepharmaceutically acceptable cations are well known to those skilled inthe art and include alkaline, alkaline earth, ammonium and quaternaryammonium cations.

The following terms, as used herein, refer to:

-   -   “halo”—all halogens, that is chloro, fluoro, bromo and iodo.    -   “C₁₋₁₀alkyl” or “alkyl”—both straight and branched chain        moieties of 1 to 10 carbon atoms, unless the chain length is        otherwise limited, including, but not limited to, methyl, ethyl,        n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl,        n-pentyl and the like.    -   “cycloalkyl” is used herein to mean cyclic moiety, preferably of        3 to 8 carbons, including but not limited to cyclopropyl,        cyclopentyl, cyclohexyl, and the like.    -   “alkenyl” is used herein at all occurrences to mean straight or        branched chain moiety of 2-10 carbon atoms, unless the chain        length is limited thereto, including, but not limited to        ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl,        2-butenyl and the like.    -   “aryl”—phenyl and naphthyl;    -   “heteroaryl” (on its own or in any combination, such as        “heteroaryloxy”, or “heteroaryl alkyl”)—a 5-10 membered aromatic        ring system in which one or more rings contain one or more        heteroatoms selected from the group consisting of N, O or S,        such as, but not limited, to pyrrole, pyrazole, furan,        thiophene, quinoline, isoquinoline, quinazolinyl, pyridine,        pyrimidine, oxazole, tetrazole, thiazole, thiadiazole, triazole,        imidazole, or benzimidazole.    -   “heterocyclic” (on its own or in any combination, such as        “heterocyclicalkyl”)—a saturated or partially unsaturated 4-10        membered ring system in which one or more rings contain one or        more heteroatoms selected from the group consisting of N, O, or        S; such as, but not limited to, pyrrolidine, piperidine,        piperazine, morpholine, tetrahydropyran, thiomorpholine, or        imidazolidine. Furthermore, sulfur may be optionally oxidized to        the sulfone or the sulfoxide.    -   “arylalkyl” or “heteroarylalkyl” or “heterocyclicalkyl” is used        herein to mean C₁₋₁₀ alkyl, as defined above, attached to an        aryl, heteroaryl or heterocyclic moiety, as also defined herein,        unless otherwise indicated.    -   “sulfinyl”—the oxide S(O) of the corresponding sulfide, the term        “thio” refers to the sulfide, and the term “sulfonyl” refers to        the fully oxidized S(O)₂ moiety.    -   “wherein two R₁ moieties (or two Y moieties) may together form a        5 or 6 membered saturated or unsaturated ring” is used herein to        mean the formation of an aromatic ring system, such as        naphthalene, or is a phenyl moiety having attached a 6 membered        partially saturated or unsaturated ring such as a C₆        cycloalkenyl, i.e. hexene, or a C₅ cycloalkenyl moiety, such as        cyclopentene.        Preferred compounds useful in the present invention include:

-   (Endo)-3-(2-methoxy-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile;

-   (Endo)-8-methyl-3-(2,2,2-triphenyl-ethyl)-8-aza-bicyclo[3.2.1]octane;

-   3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionamide;

-   3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionic    acid;

-   (Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   (Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    bromide;

-   3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propan-1-ol;

-   N-Benzyl-3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionamide;

-   (Endo)-3-(2-carbamoyl-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   1-Benzyl-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea;

-   1-Ethyl-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea;

-   N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-acetamide;

-   N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-benzamide;

-   3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-di-thiophen-2-yl-propionitrile;

-   (Endo)-3-(2-cyano-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-benzenesulfonamide;

-   [3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea;

-   N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-methanesulfonamide;    and

-   (Endo)-3-{2,2-diphenyl-3-[(1-phenyl-methanoyl)-amino]-propyl}-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    bromide.    More preferred compounds useful in the present invention include:

-   (Endo)-3-(2-methoxy-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   (Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   (Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    bromide;

-   (Endo)-3-(2-carbamoyl-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide;

-   (Endo)-3-(2-cyano-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    iodide; and

-   (Endo)-3-{2,2-diphenyl-3-[(1-phenyl-methanoyl)-amino]-propyl}-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octane    bromide.

Methods of Preparation

Preparation

The compounds of Formula (I) and Formula (II) may be obtained byapplying synthetic procedures, some of which are illustrated in theSchemes below. The synthesis provided for these Schemes is applicablefor producing compounds of Formula (I) and Formula (II) having a varietyof different R1, R2, R3 and R4 which are reacted, employing substituentswhich are suitable protected, to achieve compatibility with thereactions outlined herein. Subsequent deprotection, in those cases, thenaffords compounds of the nature generally disclosed. While some schemesare shown with compounds only of Formula (II), this is merely forillustration purpose only.

The general preparation method is shown in Scheme I. The synthesisstarted with compound 1. Reduction with lithium aluminium hydride (LAH)afforded alcohol 2. Displacement with iodine provided 3. Couplingreaction with the anion derived from HCR2(R3)(R4) then furnishedcompound 4, which was easily converted to ammonium salt 5.

A more specific preparation method leading to compounds with Formula(II) is outlined in Scheme II. Alkylation of ester HC(Ph)₂CO₂CH₃ with 3afforded compound 6. Hydrolysis of 6 generated acid 7.1,3-Dicyclohexylcarbodiimide (DCC) mediated condensation of the acidwith alcohol (R7)OH then furnished ester 8. Condensation of acid 7 withamine (R7)(R8)NH under suitable amide coupling conditions well known tothose skilled in the art such as1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCl)and 1-hydroxybenzotriazole hydrate (HOBt) provided amide 9. Reduction of6 generated alcohol 10. Reaction of 10 with acid chloride (R6)COCl oracid (R6)CO₂H afforded ester 11. Alkylation of 10 with appropriatereagents such as (R5)Br then furnished 12.

Compounds with structures similar to 6, 7, 8, 9, 10, 11 and 12 wereconverted to corresponding ammonium salts by reacting with appropriatereaction reagents such as MeBr and MeI (not shown in the scheme).Appropriate protection and deprotection methods were utilized in somepreparation procedures.

A more specific preparation method leading to compounds with Formula(II) is outlined in Scheme III. Alkylation of HC(Ph)₂CN with 3 providednitrile 13. Hydrolysis of 13 under either basic conditions (e.g., NaOHand H₂O₂) or acidic conditions (e.g., H₂SO₄) afforded amide 14.Reduction of 13 led to amine 15 that was conviently transfromed to amide16, carbamide 17, sulfonamide 18 and urea 19. Condensation of 15 withaldehyde (R8)CH(O) followed by reduction with NaBH(OAc)₃ furnished amine20 that was easily converted to amide 21, carbamide 22, urea 23 andsulfonamide 24.

Compounds with structures similar to 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23 and 24 were converted to corresponding ammonium salts by reactingwith appropriate reaction reagents such as MeBr and MeI (not shown inthe scheme). Appropriate protection and deprotection methods wereutilized in some preparation procedures.

A more specific preparation method leading to compounds with Formula(II) is outlined in Scheme IV. Alkylation of 25 with (R5)Br provided 26.Reaction of 25 with Lawesson's reagent afforded 27. Oxidation of 27 withSO₂Cl₂ and KNO₃ furnished 28 that was converted to either 29 orsulfonamide 30.

Compounds with structures similar to 26, 27, 29 and 30 were easilyconverted to the corresponding ammonium salts by reacting withappropriate reaction reagents such as MeBr and MeI (not shown in thescheme). Appropriate protection and deprotection methods were utilizedin some preparation procedures.

SYNTHETIC EXAMPLES

The following examples are provided as illustrative of the presentinvention but not limiting in any way:

Example 1

(Endo)-3-(2-methoxy-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octaneiodide

To a solution of2-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-1,1-dithiophen-2-yl-ethanol(prepared according to U.S. Pat. No. 2,800,482) (212 mg, 0.64 mmol) in 5mL methylenechloride and iodomethane (0.40 mL, 6.4 mmol), 50% aqueouspotassium hydroxide (0.25 mL, 3.2 mmol) and tetrabutylammonium chloride(5 mg, 3 mol %) was added. The reaction mixture was heated to reflux for5 d. Each day, an additional 0.2 mL iodomethane and 0.1 mL potassiumhydroxide was added. Upon completion, the reaction mixture was cooled toroom temperature, diluted with methylenechloride and washed with water.The aqueous layer was extracted with methylenechloride and the combinedorganic layers were washed with brine, dried over MgSO₄ and concentratedin vacuo. The crude product was recrystallized frommethylenechloride/ethyl acetate to give 109 mg of the title compound:LCMS (ES) m/z 362 (M)⁻.

Example 2

3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile2a) Preparation of((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-methanol

A mixture of 1,1-dimethylethyl(endo)-3-(hydroxymethyl)-8-azabicyclo[3.2.1]octane-8-carboxylate (0.50g, 2.05 mmol) and LiAlH₄ (6.16 mL, 1.0 M in THF, 6.16 mmol) was heatedat 80° C. with a microwave reactor for 60 min. The solution was thenmixed with saturated Na₂SO₄ solution, filtered through celite andconcentrated to afford the title compound (0.31 g, 97%): LCMS (ES) m/z156 (M+H)⁺; ¹H-NMR(CDCl₃) δ 1.28 (s, 1H), 1.59 (m, 4H), 1.90 (m, 1H),2.13 (m, 4H), 2.32 (s, 3H), 3.17 (s, 2H), 3.59 (d, 2H).

2b) Preparation of(endo)-3-iodomethyl-8-methyl-8-aza-bicyclo[3.2.1]octane

A solution of iodine (6.67 g, 25.8 mmol) and((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-methanol (2.0 g, 12.9mmol) in CH₂Cl₂ (120 mL) was mixed with PPh₃ (on resin, 8.6 g, 3 mmol/g,25.8 mmol). The resultant mixture was stirred for 17 hours, filtered andconcentrated to afford the title compound (2.63 g, 77%): LCMS (ES) m/z266 (M+H)⁺; ¹H-NMR(CDCl₃) δ 2.05 (m, 4H), 2.39 (m, 3H), 2.79 (d, 3H),2.98 (m, 2H), 3.45 (d, 2H), 3.81 (s, 2H).

2c) Preparation of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile

A solution of (endo)-3-iodomethyl-8-methyl-8-aza-bicyclo[3.2.1]octane(1.06 g, 4.0 mmol) and Ph₂CHCN (2.32 g, 12.0 mmol) in DMF (20 mL) wasmixed with NaH (0.288 g, 12.0 mmol). The resultant mixture was stirredat room temperature for 60 minutes. Filtration and purification via areverse phase HPLC (Gilson) then afforded the title compound (1.16 g,93%): LCMS (ES) m/z 331 (M+H)⁺; ¹H-NMR(CDCl₃) δ 1.64 (m, 2H), 2.14 (m,1H), 2.26 (m, 2H), 2.34 (m, 2H), 2.52 (m, 2H), 2.75 (m, 5H), 3.83 (s,2H), 7.39 (d, 10H).

Example 3

(Endo)-8-methyl-3-(2,2,2-triphenyl-ethyl)-8-aza-bicyclo[3.2.1]octane

A solution of triphenylmethane (0.276 g, 1.13 mmol) in THF (0.5 mL) wasmixed with n-BuLi (0.706 mL, 1.6 M in Hexane, 1.13 mmol). The solutionwas stirred for 10 minutes and added by a solution of(endo)-3-iodomethyl-8-methyl-8-aza-bicyclo[3.2.1]octane (100 mg, 0.377mmol) in DMF (1.0 mL). The mixture was stirred at room temperature for60 minutes, mixed with H₂O (0.1 mL), concentrated and filtered.Purification via a reverse phase HPLC (Gilson) then afforded the titlecompound (23.8 mg, 17%): LCMS (ES) m/z 382 (M+H)⁺; ¹H-NMR(CDCl₃) δ 1.07(d, 2H), 2.12 (m, 1H), 2.22 (m, 4H), 2.31 (m, 2H), 2.65 (d, 3H), 2.97(d, 2H), 3.63 (s, 2H), 7.21 (m, 3H), 7.30 (d, 12H).

Example 4

3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionamide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile(53 mg, 0.16 mmol) in CH₂Cl₂ (0.25 mL) was mixed with H₂SO₄ (0.28 mL,96%) and stirred at 40° C. for 30 hours. The mixture was then pouredinto ice, neutralized with NH₃.H₂O, extracted with EtOAc andconcentrated. The resultant residue was dissolved in DMSO and filtered.Purification via a reverse phase HPLC (Gilson) provided the titlecompound (17.2 mg, 30%): MS (ES) m/z 347 (M+H)⁺; ¹H-NMR(CDCl₃) δ 1.31(d, 2H), 1.98 (m, 1H), 2.28 (m, 4H), 2.39 (m, 2H), 2.67 (d, 3H), 2.79(d, 2H), 3.66 (s, 2H), 5.82 (s, br, 1H), 6.88 (s, br, 1H), 7.37 (m,10H).

Example 5

3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionicacid

A solution of2-[(3-endo)-8-methyl-8-azabicyclo[3.2.1]oct-3-yl]-1,1-diphenylethanol(100 mg, 1.56 mmol) in HCOOH (0.25 mL) was quickly added by H₂SO₄ (2.73mL, 90%) at 0° C. The reaction vial was capped immediately and stored ina refrigerator at −20° C. for 7 days. The solution was poured into ice,neutralized with NH₃.H₂O, extracted with EtOAc and concentrated. Theresultant residue was dissolved in DMSO and filtered. Purification via areverse phase HPLC (Gilson) then afforded the title compound (52 mg,48%): LCMS (ES) m/z 350 (M+H)⁺; ¹H-NMR(MeOD) δ 1.39 (d, 2H), 1.86 (m,1H), 1.97 (m, 2H), 2.30 (m, 4H), 2.69 (s, 3H), 2.84 (d, 2H), 3.69 (s,2H), 7.28 (m, 2H), 7.36 (m, 8H).

Example 6

(Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octanebromide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile(310 mg, 0.938 mmol) in acetone (6.0 mL) was mixed with MeBr (4.69 mL,2.0 M in t-BuOMe, 9.38 mmol). The resultant mixture was stirred at roomtemperature for 60 minutes and filtered. The solid washed with acetone(2×3 mL) to afford the title compound (333 mg, 83%): LCMS (ES) m/z 345(M)⁺; ¹H-NMR(MeOD) δ 1.82 (d, 2H), 2.17 (m, 1H), 2.35 (m, 2H), 2.49 (m,4H), 3.01 (d, 2H), 3.07 (s, 3H), 3.10 (s, 3H), 3.79 (s, 2H), 7.36 (m,2H), 7.43 (m, 4H), 7.49 (m, 4H).

Example 7

(Endo)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octaneiodide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile(26.5 mg, 0.080 mmol) in CH₂Cl₂ (0.5 mL) and MeCN (0.5 mL) was mixedwith MeI (0.125 mL, 2.00 mmol). The resultant mixture was stirred atroom temperature for 3 hours, diluted with DMSO (0.3 mL) andconcentrated. Purification via a reverse phase HPLC (Gilson) thenafforded the title compound (22.9 mg, 60%): LCMS (ES) m/z 345 (M)⁺;¹H-NMR(CDCl₃) δ 1.83 (d, 2H), 2.17 (m, 1H), 2.35 (m, 2H), 2.49 (m, 4H),3.01 (d, 2H), 3.07 (s, 3H), 3.10 (s, 3H), 3.79 (s, 2H), 7.36 (m, 2H),7.43 (m, 4H), 7.49 (m, 4H).

Example 8

3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propan-1-ol

A mixture of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionicacid (42.5 mg, 0.122 mmol) and LiAlH₄ (0.488 mL, 1.0 M in THF, 0.488mmol) was heated with a microwave reactor at 100° C. for 1 hour. It wasdiluted with saturated Na₂SO₄ solution, filtered through celite andconcentrated. The resultant residue was dissolved in DMSO and filtered.Purification via a reverse phase HPLC (Gilson) then afforded the titlecompound (29.1 mg, 71%): LCMS (ES) m/z 336(M+H)⁺; ¹H-NMR(CDCl₃) δ 1.40(d, 2H), 1.92 (m, 1H), 2.29 (m, 6H), 2.59 (m, 2H), 2.68 (d, 3H), 3.72(s, 2H), 4.16 (s, 2H), 7.13 (m, 3H), 7.30 (m, 7H).

Example 9

N-Benzyl-3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionamide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionicacid (82.0 mg, 0.235 mmol) in CH₂Cl₂ (3.0 mL) was mixed with PhCH₂NH₂(28.2 μL, 0.258 mmol), EDC (49.5 mg, 0.258 mmol), HOBt (3.2 mg, 0.024mmol) and (CH₃CH₂)₃N (0.232 mL, 1.65 mmol). The mixture was stirred atroom temperature for 60 hours and concentrated. The resultant residuewas dissolved in DMSO and filtered. Purification via a reverse phaseHPLC (Gilson) then afforded the title compound (29.8 mg, 30%): LCMS (ES)m/z 439(M+H)⁺; ¹H-NMR(CDCl₃) δ 1.34 (d, 2H), 1.96 (m, 1H), 2.23 (m, 2H),2.38 (m, 4H), 2.63 (d, 3H), 2.83 (d, 2H), 3.66 (s, 2H), 4.41 (d, 2H),6.93 (m, 2H), 7.22 (m, 3H), 7.38 (m, 10H).

Example 10

(Endo)-3-(2-carbamoyl-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octaneiodide

The title compound was prepared from3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionamideby following the procedure of Example 7 (33% yield): LCMS (ES) m/z 363(M)⁺; ¹H-NMR(CDCl₃) δ 1.49 (d, 2H), 1.95 (m, 1H), 2.25 (m, 2H), 2.42 (m,4H), 2.84 (d, 2H), 3.17 (s, 3H), 3.23 (s, 3H), 3.93 (s, 2H), 5.65 (s,1H), 5.91 (s, 1H), 7.39 (m, 10H).

Example 11

1-Benzyl-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea11a)3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamine

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propionitrile(250 mg, 0.758 mmol) in THF (2.5 mL) was mixed with BH₃ (2.53 mL, 1.5 Min THF, 3.79 mmol) at 0° C. The mixture was stirred at room temperaturefor 20 hours and diluted with H₂O (1.0 mL). The solution was then mixedwith K₂CO₃ (0.1 g) and stirred at room temperature for 1 hour. Organiclayers were separated and the aqueous part was extracted with EtOAc (2×3mL). The organic layers were combined, dried over Na₂SO₄ andconcentrated. Purification via a reverse phase HPLC (Gilson) affordedthe titled compound (159 mg, 63%): LCMS (ES) m/z 335 (M+H)⁺;¹H-NMR(MeOD) δ 1.35 (d, 2H), 2.01 (m, 3H), 2.34 (s, 4H), 2.55 (s, 2H),2.68 (s, 3H), 3.73 (m, 5H), 7.26 (m, 4H), 7.33 (m, 2H), 7.43 (m, 4H).

11b)1-Benzyl-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamine(50.0 mg, 0.149 mmol) in CH₂Cl₂ (2.0 mL) was mixed with PhCH₂NCO (20.4μL, 0.164 mmol) and (CH₃CH₂)₃N (62.8 μL, 0.447 mmol). The result mixturewas stirred at room temperature for 1 hours and concentrated.Purification via a reverse phase HPLC (Gilson) then afforded the titledcompound (13.0 mg, 19%): LCMS (ES) m/z 468 (M+H)⁺; ¹H-NMR(MeOD) δ 1.24(d, 2H), 1.94 (m, 3H), 2.25 (m, 4H), 2.49 (d, 2H), 2.67 (s, 3H), 3.62(s, 2H), 3.97 (s, 2H), 4.23 (s, 2H), 7.22 (m, 6H), 7.33 (m, 4H).

Example 12

1-Ethyl-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea

The title compound was prepared from3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamineand CH₃CH₂NCO by following the procedure in Example 11 (45% yield): LCMS(ES) m/z 406 (M+H)⁺; ¹H-NMR(MeOD) δ 1.03 (t, 3H), 1.33 (d, 2H), 1.94 (m,3H), 2.25 (m, 4H), 2.55 (d, 2H), 2.67 (s, 3H), 3.07 (q, 2H), 3.68 (s,2H), 3.94 (s, 2H), 7.24 (m, 6H), 7.34 (m, 4H).

Example 13

N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-acetamide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamine(33.4 mg, 0.10 mmol) in CH₂Cl₂ (0.5 mL) was mixed with Ac₂O (18.9 μL,0.20 mmol) and pyridine (16.2 μL, 0.20 mmol). The mixture was stirred atroom temperature for 1 hour and concentrated. Purification via a reversephase HPLC (Gilson) then afforded the title compound (10.7 mg, 29%):LCMS (ES) m/z 377 (M+H)⁺; ¹H-NMR(MeOD) δ 1.26 (d, 2H), 1.82 (s, 3H),1.96 (m, 3H), 2.26 (s, 4H), 2.53 (d, 2H), 2.67 (s, 3H), 3.66 (s, 2H),4.00 (s, 2H), 7.24 (m, 6H), 7.33 (m, 4H).

Example 14

N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-benzamide

The title compound was prepared from3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamineand (PhCO)₂O by following the procedure in Example 13 (8% yield): LCMS(ES) m/z 439 (M+H)⁺; ¹H-NMR(MeOD) δ 1.28 (d, 2H), 2.00 (m, 3H), 2.24 (s,4H), 2.59 (d, 2H), 2.67 (s, 3H), 3.65 (s, 2H), 4.21 (s, 2H), 7.31 (m,6H), 7.39 (m, 6H), 7.50 (m, 3H).

Example 15

3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-di-thiophen-2-yl-propionitrile

The title compound was prepared from(endo)-3-iodomethyl-8-methyl-8-aza-bicyclo[3.2.1]octane and2,2-di-thiophen-2-yl-acetonitrile by following the procedure in Example2C (34% yield): LCMS (ES) m/z 343 (M+H)⁺; ¹H-NMR(CDCl₃) δ 1.79 (m, 2H),2.21 (m, 2H), 2.33 (m, 2H), 2.62 (m, 2H), 2.73 (m, 4H), 3.80 (m, 2H),4.35 (s, 2H), 7.02 (m, 2H), 7.23 (m, 2H), 7.37 (m, 2H).

Example 16

(Endo)-3-(2-cyano-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octaneiodide

The title compound was prepared from3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-di-thiophen-2-yl-propionitrileby following the procedure in Example 7 (43%): LCMS (ES) m/z 345 (M)⁺;¹H-NMR(CDCl₃) δ 1.82 (d, 2H), 2.35 (m, 2H), 2.23 (m, 3H), 2.58 (m, 4H),2.82 (m, 2H), 3.37 (s, 6H), 4.25 (s, 2H), 7.02 (m, 2H), 7.24 (m, 2H),7.36 (m, 2H).

Example 17

N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-benzenesulfonamide

A solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamine(67.0 mg, 0.20 mmol) in CH₂Cl₂ (2.0 mL) was mixed with PhSO₂Cl (28.2 μL,0.22 mmol) and (CH₃CH₂)₃N (84.3 μL, 0.60 mmol). The result mixture wasstirred at room temperature for 1 hours and concentrated. Purificationvia a reverse phase HPLC (Gilson) then afforded the title compound (51.5mg, 54%): LCMS (ES) m/z 475 (M+H)⁺; ¹H-NMR(MeOD) δ 1.39 (d, 2H), 2.01(m, 3H), 2.30 (s, 4H), 2.69 (s, 5H), 3.60 (s, 2H), 3.68 (s, 2H), 7.12(m, 4H), 7.27 (m, 6H), 7.55 (m, 2H), 7.63 (m, 1H), 7.78 (m, 2H).

Example 18

[13-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea

To a solution of3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamine(50.0 mg, 0.149 mmol) in CH₂Cl₂ (4.0 mL), ClSO₂NCO (31.2 μL, 0.358 mmol)was added. The mixture was stirred at room temperature for 2 days andconcentrated. Purification via a reverse phase HPLC (Gilson) thenafforded the title compound (21.6 mg, 38%): LCMS (ES) m/z 378 (M+H)⁺;¹H-NMR(MeOD) δ 1.33 (d, 2H), 2.01 (m, 3H), 2.29 (s, 4H), 2.57 (m, 2H),2.68 (s, 3H), 3.69 (s, 2H), 4.01 (s, 2H), 7.25 (m, 6H), 7.34 (m, 4H).

Example 19

N-[3-((Endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-methanesulfonamide

The title compound was prepared from3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propylamineand MeSO₂Cl by following the procedure in Example 17 (28% yield): LCMS(ES) m/z 413 (M+H)⁺; ¹H-NMR(MeOD) δ 1.39 (d, 2H), 1.97 (m, 3H), 2.30 (s,4H), 2.68 (s, 3H), 2.76 (s, 3H), 3.68 (s, 2H), 3.84 (s, 2H), 7.23 (s,6H), 7.33 (s, 4H).

Example 20

(Endo)-3-{2,2-diphenyl-3-[(1-phenyl-methanoyl)-amino]-propyl}-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octanebromide

A solution ofN-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-benzamide(29 mg, 0.0683 mmol) in CH₂Cl₂ (0.5 mL) and acetone (0.5 mL) was mixedwith MeBr (0.342 mL, 2.0 M in t-butyl methyl ether, 0.683 mmol). Theresultant mixture was stirred at room temperature for 3 hours andconcentrated. Purification via a reverse phase HPLC (Gilson) thenafforded the title compound (19.6 mg, 64%): LCMS (ES) m/z 453 (M)⁺;¹H-NMR(MeOD) δ 1.20 (d, 2H), 2.32 (m, 7H), 2.65 (d, 2H), 2.98 (s, 3H),3.02 (s, 3H), 3.60 (s, 2H), 4.22 (s, 2H), 7.30 (m, 6H), 7.39 (m, 6H),7.50 (s, 3H).

BIOLOGICAL EXAMPLES

The inhibitory effects of compounds at the M₃ mAChR of the presentinvention are determined by the following in vitro and in vivo assay:

Analysis of Inhibition of Receptor Activation by Calcium Mobilization:

Stimulation of mAChRs expressed on CHO cells were analyzed by monitoringreceptor-activated calcium mobilization as previously described¹⁰. CHOcells stably expressing M₃ mAChRs were plated in 96 well blackwall/clear bottom plates. After 18 to 24 hours, media was aspirated andreplaced with 100 μl of load media (EMEM with Earl's salts, 0.1%RIA-grade BSA (Sigma, St. Louis Mo.), and 4 PM Fluo-3-acetoxymethylester fluorescent indicator dye (Fluo-3 AM, Molecular Probes, Eugene,Oreg.) and incubated 1 hr at 37° C. The dye-containing media was thenaspirated, replaced with fresh media (without Fluo-3 AM), and cells wereincubated for 10 minutes at 37° C. Cells were then washed 3 times andincubated for 10 minutes at 37° C. in 100 μl of assay buffer (0.1%gelatin (Sigma), 120 mM NaCl, 4.6 mM KCl, 1 mM KH₂ PO₄, 25 mM NaH CO₃,1.0 mM CaCl₂, 1.1 mM MgCl₂, 11 mM glucose, 20 mM HEPES (pH 7.4)). 50 μlof compound (1×10⁻¹¹-1×10⁻⁵ M final in the assay) was added and theplates were incubated for 10 min. at 37° C. Plates were then placed intoa fluorescent light intensity plate reader (FLIPR, Molecular Probes)where the dye loaded cells were exposed to excitation light (488 nm)from a 6 watt argon laser. Cells were activated by adding 50 μl ofacetylcholine (0.1-10 nM final), prepared in buffer containing 0.1% BSA,at a rate of 50 μl/sec. Calcium mobilization, monitored as change incytosolic calcium concentration, was measured as change in 566 nmemission intensity. The change in emission intensity is directly relatedto cytosolic calcium levels¹¹. The emitted fluorescence from all 96wells is measured simultaneously using a cooled CCD camera. Data pointsare collected every second. This data was then plotting and analyzedusing GraphPad PRISM software.

Methacholine-Induced Bronchoconstriction

Airway responsiveness to methacholine was determined in awake,unrestrained BalbC mice (n=6 each group). Barometric plethysmography wasused to measure enhanced pause (Penh), a unitless measure that has beenshown to correlate with the changes in airway resistance that occurduring bronchial challenge with methacholine¹². Mice were pretreatedwith 50 μl of compound (0.003-10 μg/mouse) in 50 μl of vehicle (10%DMSO) intranasally, and were then placed in the plethysmography chamber.Once in the chamber, the mice were allowed to equilibrate for 10 minbefore taking a baseline Penh measurement for 5 minutes. Mice were thenchallenged with an aerosol of methacholine (10 mg/ml) for 2 minutes.Penh was recorded continuously for 7 min starting at the inception ofthe methacholine aerosol, and continuing for 5 minutes afterward. Datafor each mouse were analyzed and plotted by using GraphPad PRISMsoftware.

1. A compound of Formula I:

wherein: the H atom indicated is in the exo position; R1⁻ represents ananion associated with the positive charge of the N atom; R2 and R3 areindependently selected from the group consisting of straight or branchedchain lower alkyl groups (having from 1 to 6 carbon atoms), cycloalkylgroups (having from 5 to 6 carbon atoms), aryl and heteroaryl; R4 is—CN.
 2. The compound according to claim 1 wherein R1⁻ is chloride,bromide, iodide, sulfate, benzene sulfonate or toluene sulfonate.
 3. Acompound according to claim 1 wherein R2 and R3 are independentlyselected from the group consisting of aryl and heteroaryl.
 4. Thecompound according to claim 3 wherein R2 and R3 are both aryl.
 5. Thecompound according to claim 3 wherein R2 and R3 are both phenyl.
 6. Thecompound according to claim 3 wherein R2 and R3 are both heteroaryl. 7.The compound according to claim 6 wherein R2 and R3 are both thienyl. 8.The compound according to claim 3 wherein R1⁻ is chloride, bromide,iodide, sulfate, benzene sulfonate or toluene sulfonate.
 9. The compoundaccording to claim 8 wherein R1⁻ is bromide or iodide.
 10. The compoundaccording to claim 1 which is:(Endo)-3-(2-cyano-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octaneiodide.
 11. A pharmaceutical composition comprising a compound accordingto claim 1 and a pharmaceutically acceptable carrier or diluent.
 12. Apharmaceutical composition for inhaled use comprising a compoundaccording to claim 1 and a pharmaceutically acceptable carrier ordiluent suitable for nasal or oral inhalation.
 13. A pharmaceuticalcomposition comprising the compound according to claim 10 and apharmaceutically acceptable carrier or diluent.
 14. A pharmaceuticalcomposition for inhaled use comprising the compound according to claim10 and a pharmaceutically acceptable carrier or diluent suitable fornasal or oral inhalation.
 15. A pharmaceutical composition comprising acompound according to claim 3 and a pharmaceutically acceptable carrieror diluent.
 16. A pharmaceutical composition for inhaled use comprisinga compound according to claim 3 and a pharmaceutically acceptablecarrier or diluent suitable for nasal or oral inhalation.
 17. Thecompound according to claim 5 wherein R1⁻ is chloride, bromide, iodide,sulfate, benzene sulfonate or toluene sulfonate.
 18. A compound

wherein: the H atom indicated is in the exo position; R1⁻ represents ananion associated with the positive charge of the N atom.
 19. Thecompound according to claim 18 wherein R1⁻ is chloride, bromide, iodide,sulfate, benzene sulfonate or toluene sulfonate.